Multiple genes relevant for the characterization, diagnosis, and manipulation of neuropathic pain

ABSTRACT

The present invention relates to the use of differentially expressed polynucleotide sequences or polypeptides for the characterization of pain status or progression, a method for characterizing pain, a method for identifying therapeutic agents for pain and the use of such sequences for the development of a medicament.

RELATED APPLICATION

[0001] This application claims priority under 35 U.S.C. §119(e) to U.S.Provisional Application No. 60/357,744, filed Feb. 14, 2002.

FIELD OF THE INVENTION

[0002] The present invention relates to the use of differentiallyexpressed polynucleotide sequences or polypeptides for thecharacterization of pain status or progression, a method forcharacterizing pain, a method for identifying therapeutic agents forpain and the use of such sequences for the development of a medicament.

DESCRIPTION OF THE RELATED ART

[0003] The effective treatment of pain requires an understanding of itsphysiology. It is well known, however, that stimuli, which activate painreceptors in one tissue, may not activate pain receptors in another. Forexample, pricking or cutting, which causes pain in skin tissue, does notcause pain in the stomach or intestine. The causes of pain in skeletalmuscle, joints, and arteries can also differ (Principles of Neurology,6^(th) ed. Adams, R. D. et al. eds. McGraw-Hill: 1997 pp. 133-134).Consequently, methods useful for relieving one type of pain are oftenless effective, or even ineffective, when applied to the alleviation ofothers. In general, neuropathic pain is persistent and is characterizedby burning, gnawing, aching, shooting, or lancinating sensations. It isfrequently associated with hyperesthesia, hyperalgesia, allodynia, andhyperpathia, and in some cases by sensory deficit or autonomicdysfunction. Unfortunately, and unlike other types of pain,neuropathic-pain tends to respond poorly to analgesic medication(Principles of Neurology, 6^(th) ed., Adams, R. D., et al. eds.McGraw-Hill 1997 p. 140).

[0004] Depending on the nerves involved, a particular instance ofneuropathic pain can be classified as a central or peripheralneuropathy. Central neuropathies arise from spinal cord, brainstem,thalamic, and cerebral damage or disease, while peripheral neuropathiesarise from damage or disease of peripheral nerves. Specific peripheralneuropathies include, but are not limited to: thoracic outletobstruction syndromes; compression and entrapment neuropathies such asulnar nerve palsy, carpal tunnel syndrome, peroneal nerve palsy, radialnerve palsy; and Guillain-Barré syndrome (The Merck Manual, 16^(th) ed.1992 1518-1522).

[0005] Neuropathic, or neurogenic, pain arises from the directstimulation of nervous tissue. Neuropathic pain encompasses a widevariety of disorders involving single and multiple nerves. Theseinclude, but are not limited to, trigeminal neuralgia and disorders dueto herpes zoster, diabetes, and trauma (including causalgia); spinalarachnoiditis and spinal cord injuries; and the thalamic pain syndromeof Dejerine-Roussy (Principles of Neurology, 6^(th) ed., Adams, R. D. etal. ed. McGraw-Hill 1997 p. 140).

[0006] Neuropathic pain is caused by a variety of factors including, butnot limited to: trauma caused by injury or surgical operation; tumors;bony hyperostosis; casts; crutches; prolonged cramped postures;hemorrhage into a nerve; exposure to cold or radiation;collagen-vascular disorders; infectious diseases such as Lyme disease,HIV; and Herpes zoster; toxins such as emetine, hexobarbital, barbital,chlorobutanol, sulfonamides, nitrofurantoin, the vinca alkaloids, heavymetals, carbon monoxide, triorthocresylphosphate, orthodinitrophenol,and other solvents and industrial poisons; autoimnune reactions;nutritional deficiency, and vitamin B deficiency in particular; andmetabolic disorders such as hypothyroidism, porphyria, sarcoidosis,amyloidosis, uremia and diabetes (The Merck Manual, 16th ed. 1992 1518).

[0007] Because so many causes of neuropathic pain exist, and because ittends to respond poorly to analgesic medication, the discovery of drugsthat safely and effectively aid in its relief has been difficult.

[0008] Whereas acute pain is generally symptomatic of tissue damage orinflammation, and lasts only as long as the underlying cause remains,chronic pain may persist indefinitely in the absence, or after recoveryof tissue pathology. It may occur as the aftermath of injury (e.g., toperipheral nerves or to the spinal cord), or in association with otherdisease states, such as herpes zoster or diabetes, but often it ariseswith no obvious cause. Chronic pain is often severe, leading to majordisability and a high suicide rate, and it is common, with a prevalenceof approximately 1% at the severe level. In contrast to acute pain,chronic pain is often unresponsive to conventional analgesic drugs(opiates and NSAIDs), and presents a difficult therapeutic problem,since its cause can usually not be resolved. Various other classes ofdrugs, (e.g. tricyclic antidepressants, some anticonvulsant drugs, suchas gabapentin, may be effective, but the therapeutic response is veryvariable and often accompanied with severe side effects.

[0009] A common feature of many clinical syndromes where chronic paindevelops is the existence of previous nerve damage, affecting peripheralnerves, the spinal cord or (as in stroke) the brain, and the concept ofneuropathic pain (i.e. pain arising as a consequence of neuronal damage)has become accepted as the underlying cause of many different chronicpain conditions seen in the clinic. Several animal models of neuropathicpain have been developed, which mimic many aspects of the clinicalcondition. These include lesions of the sciatic nerve (constriction orpartial section), constriction or section of spinal nerves, ischemiclesions of the spinal cord, induction of diabetic neuropathy, etc., andsuch models have been subjected to detailed study of the anatomical,biochemical and physiological changes that accompany the development ofthe pain state.

[0010] In the last several years a number of experimental models forneuropathic pain have been developed (Bennett & Xie 1988 Pain 33:87-107;Seltzer et al. 1990 Pain 43:205-218; Kim & Chung 1992 Pain 50:355-363;DeLco et al. 1994 Pain 56:9-16; Na et al. 1994 Neurosci Lett 177:50-52).The availability of these different models provides an opportunity toinvestigate mechanisms of neuropathic pain. Finding common features indifferent models should provide better insight into the mechanismscritical for neuropathic pain, and comparison of the models should helpto understand pain development and progression.

[0011] Kim et al. compared three of the models on basis of neuropathicpain behaviors and the effects of surgical sympathectomy (Kim et al.1997 Exp Brain Res 113:200-206). They found that the models of Bennett,Seltzer and Kim & Chung (see above) result in a very similar generalpattern and time course of evoked pain behavior, whereby the Bennettmodel showed biggest behavioral signs for ongoing pain. The same resultshave been shown for the effects of sympathectomy on the behavioral signsof evoked and ongoing neuropathic pain, respectively. Thus these threemodels can be used to discover basic common features involved inneuropathic pain.

[0012] Further animal models for pain are considered in the article ofWalker et al. 1999 Molecular Medicine Today 5:319-321, comparing modelsfor different types of pain, which are acute pain, chronic/inflammatorypain and chronic/neuropathic pain, on the basis of behavioral signs.

[0013] Object of the present invention is to identify and characterizedevelopment, conditions and progression of pain on the molecular basis.

SUMMARY OF THE INVENTION

[0014] This object is met by the use of polynucleotide sequencesselected from the group of sequences SEQ ID NO: 1 to 64 or homologues orfragments thereof or the according polypeptides for the characterizationof a) the status which elicits pain and/or b) the progression of thepathology of pain, whereby the characterization is carried out outsideof a living body.

[0015] Polynucleotide sequences SEQ ID NO: 1 to 64 are expressedsequence tags (ESTs) representing genes which are differentiallyexpressed under pain, particularly under neuropathic pain, in the modelsof Bennett, Seltzer and Kim & Chung (Bennett & Xie 1988 Pain 33:87-107,Seltzer et al. 1990 Pain 43:205-218; Kim & Chung 1992 Pain 50:355-363).

[0016] In one embodiment a combination of at least four polynucleotidesequences selected from the group of sequences SEQ ID NO: 1 to 64 orhomologues or fragments thereof or the according polypeptides is usedfor the characterization of (a) the status which elicits pain and/or (b)the progression of the pathology of pain. Such characterization iscarried out outside of a living body. The sequences are preferably ofmammalian origin.

[0017] In another embodiment a pain status is characterized by comparingof the expression of at least four genes comprising the sequencesselected of the group of SEQ ID NO. 1 to 64 or homologues or fragmentsthereof to a pain-free status. In one embodiment such characterizing ofthe pain status is performed outside of the living body. In onepreferred embodiment a pain status is characterized for assessing theefficacy of a pain treatment outside of a living body. In anotherpreferred embodiment a pain status is characterized for assessing ofanimal models for pain.

[0018] In one preferred embodiment the expression is determined from invivo samples, in vitro samples, or ex vivo samples. Such samples arederived from whole tissues, blood, cerebrospinal fluid (CSF), from cellpopulations isolated from tissues, blood or CSF or from cell lines.

[0019] In one preferred embodiment a combination of at least sixsequences is compared to non-disease status.

[0020] In one preferred embodiment the considered gene expression isincreased compared to a non-disease status. In another preferredembodiment the considered gene expression is decreased compared to anon-disease status. In yet another preferred embodiment the consideredgene expression for at least one sequence is increased and for at leastone sequence is decreased compared to a non-disease status.

[0021] In one preferred embodiment a method is provided comprising thesteps of: a) providing a test sample comprising cells or body fluidsexpressing or containing one or more genes or gene products representedby polynucleotide sequences selected from the group of SEQ ID NO. 1 to64 or homologues or fragments thereof; b) detecting expression of one ormore of the genes in the sample; c) comparing the expression of thegenes in the test sample to the expression of the same genes inreference samples whose expression stage is known, and d) identifying adifference in the expression levels of the considered sequences, ifpresent, in the test sample and the reference sample.

[0022] In another preferred embodiment, a method of identifying a testtherapeutic agent for treating pain in a subject is provided. The methodcomprises: a) providing a test cell population comprising cells capableof expressing one or more genes represented by nucleic acid sequencesselected from the group consisting of SEQ ID NO: 1 to 64 or homologuesor fragments thereof; b) contacting the test cell population with thetest therapeutic agent; c) detecting the expression of one or more ofthe genes in the test cell population; d) comparing the expression ofthe gene(s) in the test cell population to the expression of the gene(s)in a reference cell population whose disease stage is known; and e)identifying a difference in expression levels of the consideredsequences, if present, in the test cell population and the referencecell population, thereby identifying a therapeutic agent for treatingpain.

[0023] In one embodiment, the pain is neuropathic pain.

[0024] According to a preferred embodiment of the invention, geneexpression may be determined by PCR of cDNA, hybridization of a sampleDNA, or by detecting the relevant protein.

[0025] In one aspect, the method of the present invention is envisionedin the development of a medicament to treat pain.

[0026] In one embodiment an isolated nucleic acid sequence selected fromSEQ ID NO. 1 to 64 or homologues or fragments thereof or an isolatedpolypeptide encoded by one of these sequences is used as a medicament totreat pain. It is envisioned to be used alone or in a pharmaceuticalcomposition.

[0027] A kit comprising one or more reagents for detecting one or moregenes represented by nucleic acid sequences selected from the groupconsisting of SEQ ID NO. 1 to 64 or homologues or fragments thereof isalso within the scope of the present invention.

[0028] A vector comprising any nucleic acid sequence selected from thegroup consisting of SEQ ID NO. 1 to 64 or homologues or fragmentsthereof is also within the scope of the present invention.

[0029] A host cell comprising such vector is also within the scope ofthe present invention.

[0030] Antibodies that selectively bind to a polypeptide encoded by agene represented by a sequence selected from the group consisting of SEQID NO. 1 to 64 or homologues or fragments thereof are within the scopeof the present invention. As are fragments, homologues, analogues andderivatives of such antibodies.

[0031] A transgenic animal wherein at least one of the sequencescorresponding to the sequences represented by any of the SEQ ID NO: 1 to64 is altered compared to the wild type is also within the scope of thepresent invention.

BRIEF DESCRIPTION OF THE DRAWINGS

[0032] FIGS. 1 to 6 show comparison experiments of three pain models:the chronic constriction injury by the loose ligation of the sciaticnerve (CCI), the tight ligation of the partial sciatic nerve (PSL) andthe tight ligation of spinal nerves (SNL). Expression levels of 6differentially expressed sequences are represented. Each sequence iscompared over a time period of 28 days to sham operated controls. Genesare described as differentially expressed when the sequence is up- ordown-regulated at one time point in at least two of the three models.Over time the expression pattern can be determined as up-regulated inone to four time points; as down regulated in one to four time points oras mixed regulated if the type of regulation changes between up and downregulation at different time points.

[0033] X-axis describes the four time points analyzed by digitalexpression pattern display (DEPD), 1=day one post operation, 2=day 7post operation, 3=day 14 post operation, 4=day 28 post operation. TheY-axis shows Ah which represents the normalized difference of expression(peak height) of a certain transcript between a control group and atreated group. x-fold difference in gene expression is calculated by$\frac{1 + {\Delta \quad h}}{1 - {\Delta \quad h}}$

[0034] 0=no change to control, +=up regulation, −=down regulation;(0.2=1.5 fold; 0.3=1.86 fold; 0.4=2.33 fold; 0.5=3 fold).

DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENT

[0035] In the following the models are designated as follows:

[0036] The model of Bennett is based on the chronic constriction injuryby the loose ligation of the sciatic nerve. Therefore this model isdesignated as “CCI model”.

[0037] The model of Seltzer et al. is based on the tight ligation of thepartial sciatic nerve and is therefore designated as “PSL model”.

[0038] The model of Kim & Chung is based on the tight ligation of spinalnerves and is therefore designated as “SNL model”.

[0039] These designations correspond to the designations used in thecited literature.

[0040] The three models are explained in more detail in the literatureand in the examples below.

[0041] The term “polynucleotide sequence” or “nucleic acid sequence”designates in the present application any DNA or RNA sequence,independent of the length. Thus this term can describe short sequenceslike PCR primers or probes for hybridization, as well as whole genes oreDNA of these genes.

[0042] The term “polypeptide” or “amino acid sequence” designates achain of amino acids, independent from their length, however, in anycase more than one amino acid.

[0043] As “homologues” of polynucleotide sequences such polynucleotidesequences are designated which encodes the same type of protein as oneof the polynucleotide sequences described herein. Accordingly as“homologues” of a polypeptide the polypeptides are designated which havean amino acid sequence, wherein at least 70%, preferably 80%, morepreferably 90% of the amino acids are identical to one of the proteinsof the present invention and wherein the replaced amino acids preferablyare replaced by homologous amino acids. As “homologous” amino acids aredesignated which have similar features concerning hydrophobicity,charge, steric features, etc. Most preferred are amino acid sequences,containing the species- or family-dependent differences of the aminoacid sequence. Particularly as “homologues” sequences are designatedthose which correspond to one of the cited sequences in another speciesor individual. For example if in the present invention a rat model isused and the cited polynucleotide sequence encodes the rat protein, theaccording polynucleotide sequence and protein of a mouse in a mousemodel is designated as “homologue”. Further splice variants and membersof gene families are designated as homologues.

[0044] “Fragments” of a polynucleotide sequence are all polynucleotidesequences, which have at least 10 identical base pairs compared to oneof the polynucleotide sequences shown in the present application or bythe genes represented by these polynucleotide sequences. The term“fragment” encloses therefore such fragments as primers for PCR, probesfor hybridization, DNA fragments included in DNA vectors like plasmids,cosmids, BACs or viral constructs, as well as shortened splice variantsof the genes identified herein. As a fragment of a protein (polypeptide)amino acid sequences are designated which have at least three aminoacids, preferably at least 10 amino acids. Therefore fragments servingas antigens or epitopes are enclosed in this designation.

[0045] In the present application the term “sequence” is used wheneither a polynucleotide sequence (=nucleic acid sequence) or apolypeptide (=amino acid sequence) or a protein is meant. That meanswhen it is irrelevant which type of sequence is used the type is notdesignated particularly, but with the more common term “sequence”.

[0046] The basis of the models and methods described in the presentapplication is the examination and determination of the expression ofgenes, which are differentially expressed under pain or duringdevelopment of pain. Therefore for the examination each sequence can beused which allows the determination of the expression rate of theconsidered gene. Such a sequence can be at least one of thepolynucleotide sequences SEQ ID NO. 1 to 64 or homologues or fragmentsthereof, as well as the polypeptides encoded thereby, however, just aswell polynucleotide sequences and the according polypeptides can be usedwhich are (parts of) the genes represented by the polynucleotidesequences SEQ ID NO. 1 to 64.

[0047] According to the invention it has been found, that the genesrepresented by the polynucleotide sequences SEQ ID NO. 1 to 64 aredifferentially expressed correspondingly in one, two or all threedifferent models for pain, which are the above described models “CCI”,“PSL”, SNL”.

[0048] Therefore the present invention provides sequences, whichrepresent genes, which are differentially expressed under pain. Suchpolynucleotide sequences and the according polypeptides allow thedetermination and examination of pain development, conditions andprogression. These sequences have not yet been regarded in relation topain. For these examinations animal models can be used. As such a modelany animal can be used wherein the necessary preparations can be carriedout, however mammalian models are preferred. Even more preferred arerodents and lagomorpha, particularly preferred are rats, mice andrabbits. The most preferred animal model of the present invention is arat model.

[0049] The sequences of the present invention further can be used fordiagnosing a neuropathic pain status of a human outside of the livingbody by determining the expression levels of at least one of the citedsequences in comparison to the non-disease status. During treatmentperiod of a patient the expression of the presently shown sequences canalso be used for assessing the efficacy of pain treatment outside of thebody. In this case blood, cerebrospinal fluid (CSF) or tissue is removedfrom the patient and expression is determined in the samples.

[0050] For determination and comparison of the expression levels of atleast one of the genes identified in the present invention any of thecommonly known methods can be used, either on RNA/cDNA level or onprotein level. For example PCR, hybridization, micro array basedmethods, Western blot or 2-D protein gel analysis are suitable methods.One preferred method is the digital expression pattern display method(DEPD method), explained in detail in WO99/42610. The method used fordetermination of expression levels is not restrictive, as long asexpressed amounts can be quantified.

[0051] The sequences of the present invention further can be used todevelop new animal models for pain. By examination of the expressionlevels of at least one of the sequences shown it might be determined inseveral animals a procedure which is useful for generating a suitableanimal model for different interesting conditions.

[0052] In such a newly generated animal model as well as in one of theknown models the efficiency of compounds can be tested. Further asmodels for testing the efficiency of compounds assay systems can beused. Such assay systems may be in vivo, ex vivo or in vitro assays. Inany case the models are contacted with the compound(s) to be tested andsamples are obtained from these models, wherein expression levels of thesequences are determined and compared to the non-treated model.

[0053] Dependent from the used model the samples can be derived fromwhole blood, cerebrospinal fluid (CSF) or whole tissue, from cellpopulations isolated from tissue or blood or from single cellpopulations (i.e. cell lines).

[0054] In one embodiment of the invention cellular assays can be used.Preferred cells for cellular assays are eukaryotic cells, morepreferably mammalian cells. Most preferred are neuronal-like cells, likeSHSY5Y (neuroblastoma cell line), glial cell lines like TPH-1 and BV-2,astrocytic cell lines like U373MG and A7, or COS cells (African greenmonkey, kidney cells); CHO cells (Chinese hamster ovary), HEK-293 cells(human embryonic kidney).

[0055] Whereas the comparison of the expression levels(disease/non-disease status) of at least one of the provided sequencesmight give information about the examined disease status, it ispreferred to determine the expression levels of more than one of thesequences simultaneously. Thus several combinations of the sequences canbe used at different time points. By combination of several sequences aspecific expression pattern can be determined indicating and/oridentifying the conditions of the disease. The more expression rates aredetermined simultaneously, the more specific the result of theexamination might be. However, good results can also be obtained bycombination of only a few sequences. Therefore for the present inventionit is preferred to compare the expression rates of at least two of thesequences provided herein, more preferred of at least 4, further morepreferred of at least 6 of the sequences.

[0056] Since the presently provided sequences represent genes, which aredifferentially expressed, the expression rates of the single genes canbe increased or decreased independently from each other. “Independently”in this context means that the expression rate of each of the genes canbut need not be influenced by each other. In any case expression levelsdifferent from the non-disease status might be a hint to the diseasestatus, which is examined.

[0057] The disease status, which is considered in the present inventionis pain. The preferred types of pain are persistent/central pain,inflammatory pain (acute or chronic) or chronic pain. The most preferredtype is neuropathic pain. Examples of such diseases are diabeticneuropathy, post-herpetic neuralgia, trigeminal neuralgia, cancerassociated pain, spinal cord injury, multiple sclerosis, phantom pain,post-stroke pain, HIV associated pain, low back pain associatedneuropathic pain, complex regional pain syndromes, like reflexsympathetic dystrophy and causalgia, myofacial syndromes or idiopathicpain conditions.

[0058] Independent of whether a pain status is diagnosed orcharacterized, a model for pain is characterized, the efficacy of paintreatment or the efficiency of a compound in a model shall be examined,the determination of the expression levels of at least one of thesequences is carried out outside of a living body. A method to obtainsuch results comprises:

[0059] a) providing a sample comprising cells or body fluids expressingor containing one or more genes or gene products represented bypolynucleotide sequences selected from the group of SEQ ID NO. 1 to 64or homologues or fragments thereof

[0060] b) detecting expression of one or more of the genes in saidcells;

[0061] c) comparing the expression of the genes in the test cells to theexpression of the same genes in reference cells whose expression stageis known, and

[0062] d) identifying a difference in expression levels of theconsidered sequences, if present, in the test cell population and thereference cell population.

[0063] As mentioned above, detection of the expression of the genes canbe carried out by any method known in the art. The method of detectionis not limiting the invention.

[0064] Expression levels can be detected either on basis of thepolynucleotide sequences or by detecting the according polypeptide,encoded by said polynucleotide sequence.

[0065] Preferred methods for detection and determination of the geneexpression levels are PCR of cDNA, generated by reverse transcription ofexpressed mRNA, hybridization of polynucleotides (Northern, SouthernBlot systems, in situ hybridization), DNA-micro-array basedtechnologies, detection of the according peptides or proteins via, e.g.,Western Blot systems, 2-dimensional gel analysis, protein micro-arraybased technologies or quantitative assays like, e.g., ELISA tests.

[0066] The most preferred method for quantitative analysis of theexpression levels is the differential expression pattern display method(DEPD), described in detail in WO99/42610.

[0067] The sequences of the present invention can further be used foridentifying therapeutic agents and their efficiency for treating pain.For example a method can be used comprising:

[0068] a) providing a test cell population comprising cells capable ofexpressing one or more genes represented by nucleic acid sequencesselected from the group consisting of SEQ ID NO: 1 to 64 or homologuesor fragments thereof;

[0069] b) contacting said test cell population with the test therapeuticagent;

[0070] c) detecting the expression of one or more of the genes in saidtest cell population,

[0071] d) comparing the expression of the gene(s) in the test cellpopulation to the expression of the gene(s) in a reference cellpopulation whose disease stage is known; and

[0072] e) identifying a difference in expression levels of theconsidered sequences, if present, in the test cell population and thereference cell population, thereby identifying a therapeutic agent fortreating pain.

[0073] Test cells can be obtained from a subject, an animal model orcell cultures of fresh cells or cell lines. Further in vitro assays maybe used.

[0074] A method examining the different expression patterns of thedifferentially expressed genes therefore can be used for testing agentsand compounds for their efficiency for treatment of pain. Which model isused is not relevant, as long as the model allows the determination ofdifferences in expression amounts.

[0075] In such a model cells are contacted with the interesting agent orcompound and expression of at least one of the genes considered in thepresent invention is determined in comparison to the expression of thesame gene in cells which never have been contacted with the accordingagent/compound. Contacting the cells either can be effected byadministering the agent/compound to an animal or by contacting isolatedcells of tissue or blood or cells of cell lines in culture with theagent/compound.

[0076] By examination of the influence the considered agent/compound hason the expression of at least one of the genes the efficacy of theagent/compound can be estimated. This allows the decision whether it isworthwhile to develop a medicament containing such an agent or compound.

[0077] Whether the expression is determined on basis of mRNA generationor on basis of protein generation is not relevant, as long as thedifference of the expression rate can be determined. Therefore thepolynucleotide sequences, as well as the polypeptides or proteins shownin the present application can be used for the development of amedicament.

[0078] The development of a medicament can be desirable for example ifthe considered compound has an influence on the regulation of theexpression rate or on the activity of any polynucleotide sequence orpolypeptide of the present invention.

[0079] Said influence of a compound or agent can be examined by a methodcomprising contacting a sample comprising one of the nucleic acidsequences or of the polypeptides of the present invention with acompound that binds to said sequence in an amount sufficient todetermine whether said compound modulates the activity of thepolynucleotide or polypeptide sequence.

[0080] By such a method a compound or agent modulating the activity ofany of the nucleic acid sequences or any polypeptides of the presentinvention can be determined.

[0081] Furthermore the sequences itself can be used as a medicament.

[0082] An example for such a use is the use of a polynucleotide sequenceas an antisense agent. Antisense agents can hybridize to DNA or mRNA,inhibiting or decreasing transcription or translation, respectively.Thus, polynucleotide sequences of a gene, which is increased inexpression rate under pain, can be used as antisense agents to decreasethe expression rates of said gene. Further such polynucleotide sequencescan be used for gene therapy.

[0083] Another example for such a use is the use of a polypeptide or aprotein as a medicament. In case that the expression of a gene isdecreased under pain and therefore not “enough” protein is provided bythe body to maintain natural (healthy) conditions, said protein can beadministered as a medicament.

[0084] A pharmaceutical composition comprising a polynucleotide sequenceor a polypeptide according to the present invention can be anycomposition, which can serve as a pharmaceutical one. Salts or aids forstabilizing the sequences in the composition preferably are present.

[0085] For the determination of the expression of the relevant genes thegenerated sequences have to be detected. Therefore several reagents canbe used, which are for example specific radioactive or non-radioactive(e.g., biotinylated or fluorescent) probes to detect nucleic acidsequences by hybridization, primer sets for the detection of one orseveral of the nucleic acid sequences by PCR, DNA microarrays,antibodies against one of the polypeptides, or epitopes, or antibody orprotein microarrays. Such reagents can be combined in a kit, which canbe sold for carrying out any of the described methods.

[0086] Further the sequences defined in the present invention can beused to “design” new transgenic animals as models for pain. Thereforethe animals are “created” by manipulating the genes considered in thepresent application in a way that their expression in the transgenicanimal differs from the expression of the same gene in the wild type. Inwhich “direction” the gene expression has to be manipulated (up- ordownregulation) depends on the gene expression shown in the presentapplication. Methods of gene manipulation and methods for thepreparation of transgenic animals are commonly known to those skilled inthe art.

[0087] For further examinations or experiments it might be desirable toinclude any of the nucleic acids of the present invention into a vectoror a host cell. By including the sequences in a host cell for examplecellular assays can be developed, wherein the genes, polynucleotidesequences and the according proteins/polypeptides further can be used orexamined. Such vectors, host cells and cellular assays therefore shallbe considered as to fall under the scope of the present invention.

[0088] The following examples are provided for illustration and are notintended to limit the invention to the specific example provided.

EXAMPLE 1 Preparation of Rat Models

[0089] A) The CCI Model (Bennett & Xie 1988 Pain 33:87-107)

[0090] Nerve injury is created by loosely constrictive ligatures aroundthe rat sciatic nerve (4 ligatures). The ligatures evoke intraneuraledema, the swelling is opposed by the ligatures of the nervestrangulates. The constrictions remain for at least a month (hence,“chronic constriction injury”, CCI). It is known that the constrictioninjures nearly all of the nerves large myelinated axons; however, avariable but large percentage of the nerves unmyelinated axons remainedintact. Although the nerve distal to the constriction is full ofdegeneration, the nerve proximal to the constriction appears normal, andthere is no evidence of any primary afferent neuron dying. This modelhas a periphery that is innervated only by C-fibers and a greatlyreduced number of A-delta fibers, and a spinal cord that is innervatedby injured (but alive) A-beta low-threshold mechanoreceptors (AβLTMs),A-delta fibers (mostly injured, a few intact) and both injured andintact C-fiber afferents, many of which are nociceptors. This animalmodel provokes allodynia and hyperalgesia as well as spontaneous pain.Evidence of abnormal pain sensation is detected in the majority of CCIcases on the second PO day, and in nearly all cases by 5-7 days postinjury. Abnormal pain sensation appears to reach peak severity in 10-14days and to disappear in about 2 month when the pain is replaced by anapparently permanent state of hyperesthesia.

[0091] In detail, male Lewis rats (150-350 g body weight) wereanesthetized with sodium pentobarbital (50 mg/kg i.p.). Thereafter thecommon sciatic nerve on the left side was exposed at the level of themiddle of the thigh by blunt dissection through the biceps femoris.Proximal to the sciatics trifircation, about 7 mm of nerve was freedfrom adhering tissues and 4 ligatures (4.0 chromic gut) were tiedloosely around it with 1 mm spacing. The length of nerve affected was4-5 mm long. The desired degree of constriction retarded, but did notarrest, circulation through the superficial epineurial vasculature andsometimes produced a small, brief twitch in the muscle surroundingexposure. Control group animals were subjected to sham ligation (onlyleft site), which represents the same operative procedure as the ligatedone but without nerve ligation.

[0092] Four time points for gene expression profiling were chosen: 1day, 7 days, 14 days, and 28 days post operation. Animals were testedfor mechanical allodynia (von Frey test) one day before surgery andwithin 30-60 min before tissue preparation (dorsal root ganglia, spinalcord and thalamus). Tissues were frozen on liquid nitrogen prior to RNApreparation.

[0093] B) The PSL Model (Seltzer, et al. 1990 Pain 43: 205-218)

[0094] Disorders in nocifensive behavior following noxious andnon-noxious stimuli begin hours after partial sciatic nerve injury andlast for at least 7 months. This model is characterized by complexcombination of rapid onset, allodynia to touch, hyperalgesia, mirrorimage phenomena, and dependence on the sympathetic outflow. This modelresembles therefore many of the symptoms described for causalgia in man.The rapid initiation of these disorders and their contralateralappearance suggest central reorganization. This model may serve as amodel for sympathetically maintained pain (SMP).

[0095] In detail, Lewis rats (male 150-350 g body weight) wereanesthetized with sodium pentobarbital (50 mg/kg i.p.) followed byexposure of the left sciatic nerve at high-thigh level. Under 25×magnification, the dorsum of the nerve was carefully freed fromsurrounding tissues at a site near the trochanter just distal to thepoint at which the posterior biceps semitendinous (PBST) nerve branchesoff the common sciatic nerve. An 8-0 silicon-treated silk suture wasinserted into the nerve with a ⅜ curved, reversed-cutting mini-needle,and tightly ligated so that the dorsal ⅓ to ½ of the nerve thickness wastrapped in the ligature.

[0096] Control group animals were sham ligated, which represents thesame operative procedure as the ligated one but without nerve ligation.

[0097] Four time points for gene expression profiling were chosen: 1day, 7 days, 14 days, and 28 days post operation. Animals were testedfor mechanical allodynia (von Frey test) one day before surgery and30-60 min before tissue preparation (dorsal root ganglia, spinal cordand thalamus). Tissues were frozen on liquid nitrogen prior to RNApreparation.

[0098] C) The SNL Model (Kim & Chung 1992 Pain 50: 355-363)

[0099] In this model a tight ligation of the left LS and L6 spinalnerves or L5 spinal nerve alone is leading to a long-lastinghyperalgesia to noxious heat, at least 5 weeks of the affected foot.Long-lasting mechanical allodynia of the affected foot can be observedfor at least 10 weeks. This model involves a complete ligation of spinalnerves L5 and L6 or only L5 (or L4), which is more reliable to othermodels where the number and types of ligated nerves are difficult tocontrol,

[0100] In detail, male Lewis rats (150-350 g body weight) wereanesthetized with sodium pentobarbital (50 mg/kg i.p.). Thereafter thesciatic spinal nerve ligation of the spinal nerve L5 was performed onthe left site of the animals.

[0101] Control group animals were sham ligated (only left site), whichrepresents the same operative procedure as the ligated one but withoutnerve ligation.

[0102] Four time points for gene expression profiling were chosen: 1day, 7 days, 14 days, and 28 days post operation. Animals were testedfor mechanical allodynia (von Frey test) one day before surgery and30-60 min before tissue preparation (dorsal root ganglia, spinal cordand thalamus). Tissues were frozen on liquid nitrogen prior to RNApreparation.

EXAMPLE 2 Determination of Expression Levels

[0103] Gene expression profiling by DEPD-analysis starts with theisolation of 5-10 μg total RNA. In a second step, double-stranded cDNAis synthesized. Through an enzymatic digest of the cDNA with threedifferent type IIS restriction enzymes, three pools with shortDNA-fragments containing single-stranded overhangs are generated.Afterwards, specific DNA-adaptor-molecules are ligated and in twosubsequent steps 3,072 PCR reactions are performed by using 1024different unlabelled 5′ primers and a common FAM-fluorescent-labelled3′-primer in the last PCR step. Subsequently, the 3072 PCR pools areanalyzed on an automatic capillary electrophoresis sequencer.

[0104] Differential gene expression pattern of single fragments aredetermined by comparison of normalized chromatogram peaks from thecontrol groups and corresponding operated animals.

EXAMPLE 3 Sequencing and Databank Analysis of the Obtained Sequences

[0105] Differentially expressed peaks are confirmed on polyacrylamidegels by using radioactive labelled 3′ primer instead of theFAM-fluorescent primer. Differentially expressed bands are cut from thegel. After a short elution step in 60 μl 10 mM Tris pH 8, fragments arere-amplified by PCR using the same primer as used in the DEPD analysis.Resulting PCR products are treated with a mixture of Exonuclease I andshrimp alkaline phosphatase prior to direct sequencing. Sequencingreactions are performed by using DYEnamic-ET-dye terminator sequencingkit (Amersham) and subsequently analyzed by capillary electrophoresis(Megabace 1000, Amersham).

[0106] Prior to a BLAST sequence analysis (Altschul et al. 1997 NucleicAcids Res 25:3389-3402) against Genbank (1^(st) annotation, Table 1) andUnigene (2^(nd) annotation, Table 1), all sequences are quality verifiedand redundant sequences or repetitive motifs are masked.

[0107] Results are shown in Table 1. Seq Fragment ID Accession length NOnumber Name [bp] 1 novel 250 2 Y00054 HSC73 244 3 BE112971UI-R-BJ1-awa-h-11-0-UI.s1 UI-R-BJ1 155 Rattus norvegicus cDNA cloneUI-R- BJ1-awa-h-11-0-UI 3′, mRNA sequence 4 BE928419RC0-CT0499-290800-024-c01 CT0499 132 Homo sapiens cDNA, mRNA sequence 5AB030215 COXII or Elf-1 (might be 3′ similarity) 85 Rattus norvegicusmRNA for transcrip- tion factor Elf-1, complete cds. Janu- ary 2001 6BF288590 EST453181 Rat Gene Index, normal- 100 ized rat, Rattusnorvegicus cDNA Rattus norvegicus cDNA clone RGIGV25, mRNA sequence. 7BF288184 EST452775 Rat Gene Index, normal- 178 ized rat, Rattusnorvegicus cDNA Rattus norvegicus cDNA clone RGIGQ58 3′ sequence 8 novel85 9 AW534512 UI-R-BS0-ans-d-12-0-UI.s1 UI-R-BS0 167 Rattus norvegicuscDNA clone UI-R- BS0-ans-d-12-0-UI 3′, mRNA sequence. 10 M55424neurofilament NF-L 150 11 AB023781 cathepsin Y 255 12 X61479 CSF-1receptor 158 13 AJ001633 Mus musculus mRNA for annexin III 116 14 M88347Rat pseudo-cystathionine beta-synthase 325 mRNA, complete cds (type 4 offour alternatively spliced mRNAs). April 1993 15 AI058239 EST49 Ratcochlea outer hair cells 97 Lambda Zap Express Library Rattus norvegicuscDNA clone 300c 5′, mRNA sequence. March 1999 16 BF286224 EST450815 RatGene Index, normal- 76 ized rat, Rattus norvegicus cDNA Rattusnorvegicus cDNA clone RGIFN38 5′ sequence 17 BF807839RC3-CI0042-111100-011-f06 CI0042 167 Homo sapiens cDNA, mRNA sequence 18AW531794 UI-R-C4-akz-b-06-0-UI.s1 UI-R-C4 181 Rattus norvegicus cDNAclone UI-R- C4-akz-b-06-0-UI 3′, mRNA sequence. 19 novel 153 20 X83231R. norvegicus mRNA for pre-alpha- 97 inhibitor, heavy chain 3 21 M72414MAP4 138 22 V01227 alpha tubulin 209 23 AK014144 1P: Mus musculus 13days embryo head 359 cDNA, RIKEN full-length enriched li- brary, clone:3110038L02, full insert sequence. July 2001 24 AK004964 Mus musculusadult male liver cDNA, 167 RIKEN full-length enriched library, clone:1300011D16, full insert se- quence. July 2001 Weakly similar to T20253hypothetical protein F53E4.1 - Caenorhabditis elegans [C. elegans](unigene) 25 BE101201 UI-R-BJ1-aua-c-03-0-UI.s1 UI-R-BJ1 121 Rattusnorvegicus cDNA clone UI-R- BJ1-aua-c-03-0-UI 3′, mRNA sequence. June2000 Weakly similar to T21321 hypothetical protein F25B3.3 -Caenorhabditis elegans [C. elegans] (unigene) 26 AF140232 Rattusnorvegicus calcium binding 246 protein (S100A6), calcyclin 27 novel 11328 AY004290 Rattus norvegicus scg10-like-protein 254 29 AI706278UI-R-AC0-yj-c-03-0-UI.s1 UI-R-AC0 135 Rattus norvegicus cDNA clone UI-R-AC0-yj-c-03-0-UI 3′, mRNA sequence. June 1999 ubiquitin carboxy-terminalhydrolase L1 (unigene) 30 X62322 R. norvegicus mRNA for 462 epithelin 1and 2. August 1992 31 X16417 Rat mRNA for beta-globin. Septem- 208 ber1993 32 novel 211 33 AI044283 UI-R-C1-kb-f-04-0-UI.s2 UI-R-C1 190 Rattusnorvegicus cDNA clone UI-R- C1-kb-f-04-0-UI 3′, mRNA sequence. July 1999Weakly similar to T46402 hypothetical protein DKFZp434H2121.1 [H.sapiens] (unigene) 34 S74324 clone E501/543/588/5105, estrogen 82induced gene, rats, Sprague-Dawley, hypothalamus, mRNA Partial, 252 nt].April 1995 35 BF410973 UI-R-CN0-bmi-b-12-0-UI.s1 UI-R- 164 CN0 Rattusnorvegicus cDNA clone UI-R-CN0-bmi-b-12-0-UI 3′, mRNA sequence. November2000 36 novel 247 37 M31038 Rat MHC class I non-RT1.A alpha-1- 233 chainmRNA, complete cds. April 1993 38 BF288184 EST452775 Rat Gene Index,normal- 118 ized rat, Rattus norvegicus cDNA Rattus norvegicus cDNAclone RGIGQ58 3′ sequence, mRNA se- quence. November 2000 39 AI058239EST49 Rat cochlea outer hair cells 118 Lambda Zap Express Library Rattusnorvegicus cDNA clone 300c 5′, mRNA sequence. March 1999 40 D45249 RatmRNA for proteasome activator 299 rPA28 subunit alpha, complete cds.February 1999 41 novel 112 42 M17083 Rat major alpha-globin mRNA, com-336 plete cds. April 1993 43 novel 191 44 BF290323 EST454914 Rat GeneIndex, normal- 129 ized rat, Rattus norvegicus cDNA Rattus norvegicuscDNA clone RGIHV04 3′ sequence, mRNA se- quence. November 2000 45 S56463HKII = hexokinase II [rats, epididymal 230 fat pad, mRNA Partial, 266nt, seg- ment 1 of 2]. June 1993 46 novel 119 47 novel 122 48 L22643 Ratanti-acetylcholine receptor antibody 301 gene, kappa-chain, VJC region,com- plete cds. July 1996 49 AF028784 Rattus norvegicus glial fibrillaryacidic 473 proteins alpha and delta (GFAP) gene, alternatively splicedproducts, complete cds. June 1999 50 novel 216 51 BF557085UI-R-E1-go-a-09-0-UI.r1 UI-R-E1 446 HSA292757 Rattus norvegicus cDNAclone UI-R- E1-go-a-09-0-UI 5′, mRNA sequence. December 2000 Moderatelysimilar to TBB1 RAT TUBULIN BETA CHAIN [R. norvegicus] (unigene) 52AB010743 Rattus norvegicus mRNA for UCP2, 305 complete cds. February1999 53 X82396 R. norvegicus mRNA for cathepsin B. 420 September 1996 54Z12298 R. norvegicus mRNA for dermatan 205 sulfate proteoglycan-II(decorin). February 1997 55 D90035 Rattus norvegicus mRNA for 142calcineurin A alpha, complete cds. July 1999 56 D83349 Rat mRNA forshort type PB-cadherin, 150 complete cds. February 1999 57 X74125 R.norvegicus mRNA for NAD+- 298 isocitrate dehydrogenase, gamma sub- unit.July 1995 58 BF407932 UI-R-BJ0p-ait-h-01-0-UI.s2 UI-R-BJ0p 338 Rattusnorvegicus cDNA clone UI- R-BJ0p-ait-h-01-0-UI 3′, mRNA se- quence.November 2000 59 M18053 Rat ferritin heavy subunit gene 151 60 novel 30061 novel 125 62 Z22593 M. musculus fibrillarin mRNA. 204 63 AI058239EST49 Rat cochlea outer hair cells 111 Lambda Zap Express Library Rattusnorvegicus cDNA clone 300c 5′, mRNA sequence. March 1999 64 AF093567 1P:Rattus norvegicus myocilin mRNA, 150 complete cds. August 1999(Myoc/tigr)

EXAMPLE 4 Comparison of Differentially Expressed Sequences in ThreeModel Systems

[0108] Four time points for gene expression profiling were chosen forall three models: 1 day, 7 days, 14 days, and 28 days post operation.Behavioral testing of the animals was performed one day before operationand 30-60 min prior to the tissue preparation. After DEPD analysisdifferentially expressed peaks obtained for the three different painmodels were compared to identify overlapping gene expression patterns.

[0109] Results are shown in Table 2. For 6 selected sequences theregulation is further shown in FIGS. 1 to 6 (Regulation of Seq No: 1,18, 20, 22, 30 and 59). Seq ID No. Regulation 1 up 2 up 3 up 4 down 5mixed 6 up 7 down 8 up 9 mixed 10 down 11 up 12 up 13 up 14 up 15 down16 mixed 17 up 18 down 19 down 20 up 21 up 22 mixed 23 down 24 down 25up 26 mixed 27 mixed 28 mixed 29 mixed 30 up 31 mixed 32 mixed 33 up 34up 35 up 36 up 37 up 38 down 39 mixed 40 up 41 down 42 up 43 mixed 44mixed 45 up 46 down 47 up 48 up 49 up 50 down 51 mixed 52 up 53 up 54mixed 55 up 56 mixed 57 mixed, 58 down 59 up 60 up 61 up 62 up 63 mixed64 up

[0110] For each DNA fragment, gene expression patterns obtained in thethree pain models were matched. “Up”, “down”, and “mixed” regulation isdefined as overlapping pattern in at least two of the three models atone or more time points.

1 64 1 193 DNA Rattus norvegicus 1 acgcgagaga ctcagtacca actcctcaactaaccttata atgactgcat ggatgccccc 60 caccctatca cacattcgaa gaaccttcctacgtaaaagt taaataagaa aggaaggatt 120 cgaaccccct acaactggtt tcaagccaatttcataacca ttatgtcttt ctcaataaaa 180 aaaaaaaaaa aaa 193 2 244 DNA Rattusnorvegicus 2 cttattgatt gggagtgacc atatgatgca agctgtacag agtgctggtggcgtgcctgg 60 aggaatgctg gtggctgccc tgggtggagg agctcctcca tctggatggtgcttcttcag 120 gccccaccat tgaagaggtc gattaagtca aagtagaggg tatagcattgttccacaggg 180 acccaaaaca agtaacatgg aataataaaa ctatttaaat tggcaccaaaaaaaaaaaaa 240 aaaa 244 3 155 DNA Rattus norvegicus 3 tccatatcgcatgatcctcg cttaagttag ggattgagca cgacgaagca ttttagcgat 60 tcggatggtcgcacttgact atgtagtttc cagtgcacag agcgacctaa tggctcaata 120 aacttggcctcaacagtcaa aaaaaaaaaa aaaaa 155 4 132 DNA Rattus norvegicus misc_feature(1)...(132) n = A,T,C or G 4 gcggttgttn tcggggaggg ctagcgacaa ctgactgatctattaaacaa agcatccgat 60 cgtccgcggt cgggtgttga cgcgatgtga tttctagcccagtgctctag aatgtcaaag 120 aaaaaaaaaa aa 132 5 85 DNA Rattus norvegicusmisc_feature (1)...(85) n = A,T,C or G 5 gtcnaccatt gtactagaaactagtgctct aaaatatttc gaaaactgat cagcttctat 60 aaatttaaac caaaaaaaaaaaaaa 85 6 100 DNA Rattus norvegicus misc_feature (1)...(100) n = A,T,Cor G 6 agaacaggtg agntgtcggc agcacgcggc atacgattac ccacactaat tattttcggc60 gtaaaacgtg ccaactataa atctcaaaaa aaaaaaaaaa 100 7 178 DNA Rattusnorvegicus 7 cctgattgtg atgagctgta ctgttcgtta gttagcgact attagtcttacttaatctgt 60 acttctgatc tggagcattc gaggtcggtt tctatctatt gtacaatttctgccctagta 120 cgtacggacc cgagaaatgg agcctcctat accataattg ctcccaaccaaaaaaaaa 178 8 85 DNA Rattus norvegicus misc_feature (1)...(85) n =A,T,C or G 8 tggcagcacg nggacatacg anntacccan attaattatt tactggcgtaaacgtgccaa 60 ctatacatct caacaaaaaa aaaaa 85 9 167 DNA Rattus norvegicus9 ggctcatttt ctcacaccca cgtgacacac gtcatacgtt tgtgtatact gtgcaggcaa 60atgctcactc tccccggaaa gagaattttg aatgtggaaa ggagaaaggc agaggaggaa 120gaaaattctt ttaatttaaa gcacacattc aaaaaaaaaa aaaaaaa 167 10 150 DNARattus norvegicus 10 caaatcagtc gtcctctact cctccatccc ttctccccctccattccgct ctagctatgt 60 gaaacttcag tggttagaaa ttaaagacct cgcagttatgtgcaataaat agtaaataca 120 agttacagtg gatgacaaaa aaaaaaaaaa 150 11 255DNA Rattus norvegicus 11 gggaggagcg accgacacga actagaaatt atgagcacaagtacactgga atcctccagc 60 ttcagagctg cttcctccac ccacagacct gcttcctcctccaccgtgcc cgagcaggcc 120 taacctccag accgtcagag aggacagcta tggtctaggacagttctggt gttaccctgg 180 agtccacggg aggggaacta gtccagactg cctgagatgagtaaagtatc tggcgtcacc 240 aaaaaaaaaa aaaaa 255 12 158 DNA Rattusnorvegicus misc_feature (1)...(158) n = A,T,C or G 12 acacccanaccgtagacaat acctacctnc cacagcctac ttgtcctgtc tctagctact 60 actccacgcagatactgcgt gctcctctcc aaactgactc gtcctcatta acagtcaaca 120 ttaaactaacagcattaaca caaaaaaaaa aaaaaaaa 158 13 116 DNA Rattus norvegicus 13caggcgctgt ctaagttact ttgtattgtt ttccgtacta ctaatactgt atgttgcctg 60gtgccaacaa atacttcaaa atacatttgt tgtaaatgca aaaaaaaaaa aaaaaa 116 14 325DNA Rattus norvegicus 14 gcgccaagca tccgtgctag atagtgtctc gcctagagacggccattcct gagaaaggga 60 gaccgggact gatcgttctc atcctcaggg gcagagttggccccaccacg ggccatgtgg 120 gttctaaatg agggtagtgt tccagtgacc tgagacgccacagctgtgag ctccacgtcg 180 tgccggtacg cgactgacac cgacctgggt catgaccctgcttcgcagtt cctcctcaca 240 ttatcctcct ttgccgacac gcacctacta tcggtctcaactcttcttat aaatgattca 300 catacctgtg aaaaaaaaaa aaaaa 325 15 97 DNARattus norvegicus 15 gtggtttatc tatttacaat ttctcccagt ttcgaaaggacaagagaaat ggagcctcct 60 taccataagt gctcccaacc aaaaaaaaaa aaaaaaa 97 1676 DNA Rattus norvegicus misc_feature (1)...(76) n = A,T,C or G 16ctgggcacna nccnanaatg taacttacct ataacctnaa agagggacag ctctttagga 60aaaacaaaaa aaaaaa 76 17 167 DNA Rattus norvegicus misc_feature(1)...(167) n = A,T,C or G 17 cgtntgatgc gngacnacga gggcggcaggggcntnccna tcgtgccgct anatggtaat 60 gaccgtggct ggtacaacga cggaccagagcgacagcatt cgccaggaat gtttaacatt 120 acatcnaaaa aaancaaaan acgctcnccacnaacaccna aaaaaaa 167 18 181 DNA Rattus norvegicus misc_feature(1)...(181) n = A,T,C or G 18 atgaaattat agagatgcgt cgatctatagcgtgagttga tggactactg tctggcaggt 60 catgttacgg attgacgcat accacagtgtgagaaactcc aaggatatca gacacgtagg 120 agtcaagcac aaagcccgat attctaatttacttaagaat gtccaanaaa aaaaaaaaaa 180 a 181 19 153 DNA Rattus norvegicusmisc_feature (1)...(153) n = A,T,C or G 19 gaaatgatgg gttgcacccccagatngttt anagacccat ctgccataga ngtacngnaa 60 actantagta ccatntctntcaaataaaga ataancnggt aggcgaagcg atgnnnacaa 120 nacacacnaa cnaaaacaaaaanaaaacgc aaa 153 20 97 DNA Rattus norvegicus 20 ttgtgctgca gtagcaaggcctggaacagg acacagaatg ggctcgtgtg gaaggactgg 60 gacaataaag tgggtaaaccaaaaaaaaaa aaaaaaa 97 21 138 DNA Rattus norvegicus 21 gcacctgaggagggacggcg cactctgacc accctccggg actgtatcac cacaagattc 60 ttatagcactagtattttct gtattaaagt ttgcatggtt tctaataaag aattcaaacc 120 taaaaaaaaaaaaaaaaa 138 22 209 DNA Rattus norvegicus misc_feature (1)...(209) n =A,T,C or G 22 gctggggcgg attacctagt taattttctc tggtcaccca tacacagtacgtgggctcga 60 tcttttaatt ttgtatagtn gcactgtgtg cgtttcatac agtgtggctngactgaaagt 120 tgtgaatgat ttgtcaggag acccgagacc gtccagttca ctgatgggtttnaaataaaa 180 tactccctga tcttaaaaaa aaaaaaaaa 209 23 359 DNA Rattusnorvegicus 23 cctggtgtga ctgaggcatg ggcagtggac ggatgcccat gccgtgctttgcgccaggct 60 actgtgaatg gtggaagcag gagggagagc cacctggtac tttcgtccctgcctcctcct 120 ccgttccgga tctctcccct ctgtccaggg gtgtgtgacg tttctcctgcatgtttttac 180 tgatgttcgt gctggccgcc ctcagcccgg agcctgggag aggctttggtgcctcgcatc 240 agacttcggt gctccgtggc ggtgaagccc ttaaatgctt tgtatattttctctattaga 300 tctcttttca gaagtgtctg tagaagatta aaaaaaacaa aaacaaaaaaaaaaaaaaa 359 24 167 DNA Rattus norvegicus 24 cgtgggagcg tgaggagcgtagactgtggc gtgaagctcc gatcccaatt cgcggtggtg 60 gggaccaaca gactagcttcatcctgtcct tgtagctgta gctgtacccc tgaagctcag 120 ccagctctga ctttataaaacataaaacct caaaaaaaaa aaaaaaa 167 25 121 DNA Rattus norvegicusmisc_feature (1)...(121) n = A,T,C or G 25 aagacagaac gacaacnaggaccgtagaag cgtgtgggtt gggcggagcc tacgggttct 60 acgcatgcta caccatatgtgcttagtaaa tgtctgttca atcgaaaaaa aaaaaaaaaa 120 a 121 26 245 DNA Rattusnorvegicus 26 gttggcagct gcaggatctg aaattgcatg ctgatggatg atctggaccgtaacaaggat 60 caggaagtaa acttccagga gtatgttgcc ttcctggggg ccttggctttgatctacaat 120 gaagctctga aataaaatgg gaaggcagag atgccttctg ggaggctatcccagtcaagt 180 ccagtggggg ggtaattata caataaatat ttcgtttttg ttatgtctaaaaaaaaaaaa 240 aaaaa 245 27 113 DNA Rattus norvegicus misc_feature(1)...(113) n = A,T,C or G 27 agacgncgac gttcagtnac tacgatgggacggattcgtc atgcccctac aatctggttt 60 caagcccctt tcactaacca ttatgtctttcgcaancaca aaaaaaaaaa aaa 113 28 254 DNA Rattus norvegicus misc_feature(1)...(254) n = A,T,C or G 28 gtccanccna cnatatgact tgtgccgctcgcatctgctn tcgtacccca gcttccngct 60 tttctcccac atactagaac tgtccagtcctatgtagatg tagtctgacc taggactcta 120 tcctgaagga gctccctagg caggaatatggtcccctatt cagacactag gccaggtgtg 180 actggggctc tctnagtggc cctcttagtgnatgtgttgg caaccttaat aaatctagtg 240 gcagtggcaa aaaa 254 29 135 DNARattus norvegicus 29 tgacagaggc agcctacctc gtgtctgtac tgctctctatggtctctttg gttctgtaag 60 tgacggcctg gatgtggttg tctagtcctc agaggaagaataaaactttg ctgctggcaa 120 aaaaaaaaaa aaaaa 135 30 462 DNA Rattusnorvegicus 30 aggtgacggc accaactgct tatacggact gcaggcgtct gatccagccttccattgtac 60 ctgaactttg gctctaaggt tgggaatgtg gaatagtggt gccggacatttctgccatga 120 taaccagtcc tgttgtaaag acagcgcaag ggaggctggg cctgctgtccctatgtaaag 180 ggtgtctgct gtagagatgg acgtcactgt agtcccattg gcttccactgttcagccaag 240 ggaaccaagt gtttgcggaa gaagacccct cgctgggaca tacttttgagggatccagcc 300 ccaagaccgc tactgtgagg aagggctaac gactaaagaa ctccacagtcctgggaaccc 360 tgttctgagg gtatccacca ctcaggcctc cctggcacct cttcctttagtctccccggc 420 ctactcattc tgagtcaccc catcaccatg gaaggtgggg cc 462 31 208DNA Rattus norvegicus 31 gggtaggcgc ttcagaaagg tggtggctgg agtggcagtgccctggctca caagtaccac 60 taaacctctt ttcctgctct tgtcttgtgc aatggtcaattgttcccaag agagcatctg 120 tcagttgttg tcaaaatgac aaagaccttt gaaaatctgtcctactaata aaaggcattt 180 actttcactg caaaaaaaaa aaaaaaaa 208 32 211 DNARattus norvegicus misc_feature (1)...(211) n = A,T,C or G 32 gggtagngngaagaaacgng antnaccgtc ttanaactcg catttacgct caacacnctt 60 ntncccgacgcgagctttct natgcaagag gttatgnttc cggccggatc atctaggcca 120 aatggtgtgcaagactgagn aagggaagga agggagaaaa gcgcgtactc ctactttaga 180 gagtagctggttgcccngca aaaaaaaaaa a 211 33 190 DNA Rattus norvegicus 33 cagccagcggctaggcgccg acaccccctg acggcttcgc agataagtta ctatttaaca 60 acgatttaacaggcaactag aactctaaac ttaatagagt agcttactag agataataat 120 attttactattaagatttta aaagacacag tatggaggtt tatttaaaca atttgaaaaa 180 aaaaaaaaaa190 34 82 DNA Rattus norvegicus misc_feature (1)...(82) n = A,T,C or G34 agagagacna ggtagtctcc cttactagcc ttcancttat ataaaaacaa cctaatgggc 60taaaacaata aaaaaaaaaa aa 82 35 167 DNA Rattus norvegicus 35 agactttgctctgtgtcagc atgtaagtac tcactcctga tgcagtcctc catcgctgtt 60 gggaaagaggaatctatggc atagagcctg tgcttatcac taacattttg aaatttccaa 120 accctttagttaaataaatc tcttcttcct aaaaaaaaaa aaaaaaa 167 36 247 DNA Rattusnorvegicus misc_feature (1)...(247) n = A,T,C or G 36 gagagagcnactccanattc tncaccttct ttnttnntat ttagggccca ncctgcagtc 60 ncttctctctgnattctgca aagctgcagg ggctgaggat tcgagcacgt ctcggnggag 120 gnggctggctccgtagcggc actgatnact gatgtatcta ttgagcacgg tccaatnatt 180 atctttangtatgcagctgg gaataaagta gaccccatgt gttagggggt ccgaaaacaa 240 aaaaaac 24737 233 DNA Rattus norvegicus 37 tgtgagcctc cgacgcttta gctcaccccctcactctaca ctgagaataa gaatctgagt 60 gtgaacttga ttgttacaca taccttgacacaagtgtgga tagcttttca aattactgga 120 tggaatactt agagcgtttg ttgttgttggggtattagct attgttcttg tatttgtcct 180 ttgaaaacaa ataaactggc acgatgaacgaagccttcca acaaaaacaa aaa 233 38 118 DNA Rattus norvegicus 38 gggcgacggacatcaggcgg tcctatctat ttacatttct cccagtacga aaggacaaga 60 gaaatggagcctccttacca taagtgctcc caaccaattt atgaaaaaaa aaaaaaaa 118 39 118 DNARattus norvegicus 39 agagagacta cggagcatca ggtcggttct atctatttacaatttctccc agtacgaaag 60 gacaagagaa atggagcctc cttaccataa gtgctcccaaccaaaaaaaa aaaaaaaa 118 40 299 DNA Rattus norvegicus 40 aacgcgtgcagcggagtcac ggagaccggc tgatggcatg gagatcgtaa cgcttatgct 60 gtgttatatgacatcatcct gaagaacttt gagaagctca agaagccccg tggagagaca 120 agaggggatgatctattgag ccccctctct cgtactatga tgggtataac agaaaccttc 180 ctactcttgactaggaactc tagactgcac ccagtcttct tcctgctggg gtctcctccc 240 tcactctgccttccaaacac aataaataca tagttgccca ccaaaaaaaa aaaaaaaaa 299 41 112 DNARattus norvegicus misc_feature (1)...(112) n = A,T,C or G 41 aagttttagtttaaccaata tttagtcagc tattcagttg caatcaatct ntatacgaga 60 ttcattctaacaaatntctc tcatgtacca cacacacana aaaaaaaaaa aa 112 42 336 DNA Rattusnorvegicus 42 gtgcggcaag tgagacacgt gataactgct gtgtgcctgt ccactctgagcgacctgcat 60 gcccacatac tgcgtgtgga tcctgtcaac ttcaagttcc tgagccactgcctgctggtg 120 accttggctt gccaccaccc tggagatttc acacccgcca tgcacgcctctctggacaaa 180 ttccttgcct ctgtgagcac tgtgctgacc tccaagtacc gttaagccgcctcctgccgg 240 gcttgccttc tgaccaggcc cttcttccct cccttgcacc tatacctcttggtctttgaa 300 taaagcctga gtaggaagca aaaaaaaaaa aaaaaa 336 43 295 DNARattus norvegicus misc_feature (1)...(295) n = A,T,C or G 43 ggtcgtattctatatcagat gcttacacca catgaaatac agtctcctct ataggctcat 60 tcatctcacttacggccgtc cttgtaatga tcttcatgat ttgagaagcc ttcgcatcaa 120 aacgagaagtactctcaatt tcctactcct caatataacc ctagaatgac tgcatggatg 180 ccccccaccctaccacacat tcgaagaacc ttcctacgta aaagttaaat aagaaaggaa 240 ggattcgaaccccctacaac tggggttntc cagagggcgc gaaattntct cacta 295 44 129 DNA Rattusnorvegicus 44 45 230 DNA Rattus norvegicus 45 aagccgggac accaagctaagtgctatccg cttgctagct gtagcttgag catcctcctg 60 tatctctgat aatgtgcgatctccgagagg aacagaactg tcatataagc gaagttgagc 120 cttactgatc ccgtgggcgaagttgcgacg ggacgctgag caactagacc ggtcggcagg 180 agtgagactt aggtgccttctagatagttg tgacttaaaa aaaaaaaaaa 230 46 119 DNA Rattus norvegicusmisc_feature (1)...(119) n = A,T,C or G 46 cctaacanaa natagaggcagaggcnggag gattctgagt tgaaggcagc ctggtctaca 60 aagtgagttt caggactgccaggggctatc tcaaaacaaa aaaccaaaaa acaaaacaa 119 47 122 DNA Rattusnorvegicus misc_feature (1)...(122) n = A,T,C or G 47 cttaancagtgtataagagg cagaggcngg aggtntctga gttgaaggcc agcctggtct 60 acaaagtgagtttcaggact gccagggcta tctcaaaaca aaaaaccaaa aaaaaaaaaa 120 aa 122 48 301DNA Rattus norvegicus 48 atcgtcgaca taatcttacc ccctatcttt tcagcttgtcatagacatca tcctcatccg 60 tcgtcaagag cttcaacagg aatgagtgtt agacccaaaggtcctgaggt gccacctgct 120 ccgccagctc cttccaatct tccctcctaa ggtcttggagacttccccac aagcgaccta 180 ccactgttgc ggtgctccaa acctcctccc cacctcatcctccttccttt ccttggcttt 240 gatcatgcta atatttgggg aatattaaat aaagtgaatctttgcacttg aaaaaaaaaa 300 a 301 49 473 DNA Rattus norvegicus 49accaagattt atggcgatac ttgtgacaat gcgagttggt tagttgtagg caactagtta 60cacttggctc tgaatccttg gactcacggc aatgacctgt aactctacaa gagacactga 120aacagtgaga gagggacttc cataccactg ggcaggctac aggcgcgtct cagttgtgac 180ggtctattcc tggttgctca gtccccaacc tcgcgtcacc ctgggaccgc cgattctcaa 240cctgaaagag tccacaacca tccttctgag tgccctccat ccccacaaac cactagcatg 300ttgtactcca agcccaaggg gccccattcc ctttcttatg ccatgtcacg gagtatcagg 360ctcagcactt caagcgtcca tcctggtttg aacagtttgg gcaaactgac accacgtagt 420gtaacaccca tgcctggttg tgcagtgcca tccgtcattc gtgtaccctc act 473 50 216DNA Rattus norvegicus misc_feature (1)...(216) n = A,T,C or G 50aggaatacat gcngtactcn attttttgtt nngtttcttt atctgatggn catgattaac 60gagtggacgg ccgggagnat nccgtattag cgccgttagg caggtagaac attcttgtgg 120acccggcgca atagacggac caagactcga acaggccctt acgccaacgg aaactggttg 180tccaccttaa ggtcaacaaa aacacacaca cacccc 216 51 446 DNA Rattus norvegicus51 gctgtctgtg acattccacc tcggggccta aaaatgtccg ccaccttcat tggcaacagc 60acggctattc aggagctgtt caaacgcatc tcagagcagt tcacagccat gttccgacgc 120aaggccttcc tacactggta cacgggcgag ggcatggatg agatggagtt cactgaggct 180gagagcaaca tgaatgacct ggtgtccgag taccagcagt accaggatgc tacagctgag 240gaggaaggag agtttgagga ggaggctgag gaggaggtgg cttagagctg tcttagtcac 300tacagcatgg gagcagtgtg aactctttat tcattcacag cttgtctgct agccatgtcc 360actgtgcatt tgctgtcctg tgtcctgaca tcacttgtac agataccacc attaaagcaa 420ttcatagtgt caaaaaaaaa aaaaaa 446 52 276 DNA Rattus norvegicus 52gcgcagtacc tagggataca gcgcatccta tttaagagtt catatcgaca attagggttt 60acgacctcga tgttggatca ggacatccca atggtgcaga agctattaat ggttcgtttg 120ttcaacgatt aaagtcctac gtgatctgag ttcagaccgg agcaatccag gtcggtttct 180atctatttac aatttctccc agtacgaaag gacaagagaa atggagcctc cttaccataa 240gtgctcccaa ccaatttatg aaaaaaaaaa aaaaaa 276 53 420 DNA Rattus norvegicus53 ggatggccct gtaagctagt cacttctagg actgtggcag gctttgatgg cccaatgcag 60ttctctggag aaaactacct ttccccaagg catctgcacc cattgacaat ggtaatgtgc 120ccatctctcc ttggtcctgc cctcaaccga tgcttttcca gtcagggttt tgttttttgt 180tttgttttgt gtacctcaac tgagttatga agatttgtac tggttttaca gatcatctca 240tcgtatggat tagaacaagc ttcgtggtca gtttgctggg tgaccggcag acaccacaat 300caaactagtc tgggaaaaac ctgctttttt gttgtaggtg ccacgtaacc ctgtcagttt 360aacaaggaat gaccgtgcca ataaaccaat tctccctctg cttgaaaaaa aaaaaaaaaa 420 54205 DNA Rattus norvegicus 54 accccaacag actagagact ctaccgttcg tagtttatagcatctctctc cggtattggc 60 aattcaccca cacaccttca gatgtgtctt cgggcgctctaccattcaac ttgggaacta 120 cgagtaactc ccacacagcc tcatattata atcgggaacaaaaaaaccaa tctgtcaata 180 ttattgctaa aaaaaaaaaa aaaaa 205 55 142 DNARattus norvegicus 55 aacagccaac catctaaaca tgaaaacaac gatgtcaaactaaataagga ctatatactt 60 gataatgttt tgctactctt gtcagacaat ggctataaactgaattaggc agtcttaaca 120 aaaaataaaa aaaaaaaaaa aa 142 56 150 DNA Rattusnorvegicus misc_feature (1)...(150) n = A,T,C or G 56 gacgtggnaacagccngggg tggtggtggc cacgtctggt agcatgtgct gacctcacct 60 cttagccacggtgtgacctt aagacaagtc atttagcttc caagtttcag ttccttgcag 120 cacctagatgacatttaaaa aaaaaaaaaa 150 57 298 DNA Rattus norvegicus 57 gatagacgaagcacactgcc tactcctatg cttgttcact ccatcgaaaa gtgatgaaac 60 ctgatggagaatgaacatat gcatacccca gacattggag gccaggggaa cacataccaa 120 ggcatccaggatatgattcg tcacctgcgc atgattaatg gatgggctgt ggaggtgtaa 180 ctatgcctgcagttagctca gtctgactga acgactcgct tctcatatta gcacaccagc 240 tagcttaggggacaggctcc agaataaagc cacttgtgtt ccaaaaaaaa aaaaaaaa 298 58 338 DNARattus norvegicus misc_feature (1)...(338) n = A,T,C or G 58 gactagtcaggtctcaacat ctacntgaca tggctggcac tgtgtgtatg tggtgcacaa 60 acacacgaaggcagaacacc taaaaggggt atatgtgtta tcatttaagt gtctctttaa 120 atgaaaagccttcaaccagg atttccctcc ttacaatata gatttgatgt ccaccctgtg 180 tcatgggaactgagaggaag ggcagtataa atctgagagg ttcctttgtg tggtggaccc 240 cgaagaagaaagccccatgg ctgaacagct gttgtctcct cctaccccac agctttccct 300 aataaagggattgttatttt gaaaaaaaaa aaaaaaaa 338 59 108 DNA Rattus norvegicusmisc_feature (1)...(108) n = A,T,C or G 59 ggaggcgncg tnantttctatagtgcacta aacatctgct taaaagttct ttaattgggt 60 accatttctt caaataaagaatttgggtac ccaaaaaaaa aaaaaaaa 108 60 300 DNA Rattus norvegicusmisc_feature (1)...(300) n = A,T,C or G 60 gaatatccag cacaaacaactacngntttt accnnggggg acctgttcct cacatcagct 60 tagatatcta gtaagatccgtagntatata ctatgtgata cagcagttac acttaaggga 120 acnntgaaag ccgagcatggcaggtgnact ctcgattcgg tgatcaatgg tttcggtagn 180 gacatacatg tcgagngccgcgatgacagt tatatatgct cantcgcgtg gcatcgangt 240 attatagcaa tataggatacgataaanatg tcgccngcaa caccaaaaaa aaaaaacaaa 300 61 125 DNA Rattusnorvegicus misc_feature (1)...(125) n = A,T,C or G 61 ctaaccagcnatatacgagg cngaggccgg cggatctctg agttagaagg ccagcctggt 60 ctacaacagtgagtnttcag gactgccagg ctatctacaa aacaaaaaac caaaaaaaaa 120 aacna 125 62204 DNA Rattus norvegicus misc_feature (1)...(204) n = A,T,C or G 62taccagacag ttgaactata gccttatgaa tttagacacg ctgctngttg taggtgtgta 60caggccacct cccaaggcga ataactgaaa ctcgactgnc tggattgaag aaatgtgtgt 120gttgctactg ttgcacatgt gtgcttatga cttgttgcag agggatttcc tattaaaaga 180ctaatctgtc aaaaaaaaaa aaaa 204 63 111 DNA Rattus norvegicus misc_feature(1)...(111) n = A,T,C or G 63 gacgggagca ncagggnggg ttnnattctattnacgnttt tctcccagta cgaaaggaca 60 agagaaatgg agcctcctta ccataagtgctcccaaccaa aaaaagaaaa a 111 64 150 DNA Rattus norvegicus 64 gagaatacgggagggggtga caccactcct gtggttagaa ctgctcctgc agtatttact 60 ccagtttctaaggctacaga tatagcatgt acattaaacc ctttgccctg tgaaataaag 120 ttatcttacacgaaaaaaaa aaaaaaaaag 150

What is claimed is:
 1. A method for characterizing a pain status,comprising: a) detecting a level of expression of at least four genesequences selected from the group consisting of SEQ ID NO. 1 to 64 orhomologues or fragments thereof, in a test sample from an individualwith pain; b) detecting a level of expression of said at least four genesequences in a control sample; and c) comparing the levels of expressionin the test and control samples, thereby characterizing the pain status.2. The method of claim 1, wherein the control sample is from anindividual without pain.
 3. The method of claim 1, wherein the controlsample is derived from a cell line.
 4. The method of claim 1, whereinsaid individual is an animal.
 5. The method of claim 1, wherein saidindividual is a human.
 6. The method of claim 2, wherein said individualis an animal.
 7. The method of claim 2, wherein said individual is ahuman.
 8. The method of claim 1, wherein the test sample and controlsample are derived independently from a source selected from the groupconsisting of whole tissues, blood, cerebrospinal fluid, isolated cells,and cell lines.
 9. The method of claim 1, wherein the test sample andcontrol sample are derived independently from an in vivo sample, an invitro sample, or an ex vivo sample.
 10. The method of claim 1, whereinthe levels of expression in the test sample are increased relative tothe levels of expression in the control sample.
 11. The method of claim1, wherein the levels of expression in the test sample are decreasedrelative to the levels of expression in the control sample.
 12. Themethod of claim 1, further comprising detecting a level of expression ofsaid at least four gene sequences in the test sample prior to andfollowing administration of a pain treatment; and comparing the levelsof expression prior to and following administration, thereby assessingthe efficacy of the pain treatment.
 13. The method of claim 1, whereinsaid individual is an animal used in an animal model for studying pain.14. The method of claim 13, wherein the animal is subjected to a painstimulus.
 15. The method of claim 14, wherein the animal is administereda compound that may alter the pain status.
 16. The method of claim 1,further comprising detecting at least six gene sequences selected fromthe group consisting of SEQ ID NO. 1 to 64 or homologues or fragmentsthereof, in the test sample and the control sample.
 17. The method ofclaim 1, further comprising detecting at least eight gene sequencesselected from the group consisting of SEQ ID NO. 1 to 64 or homologuesor fragments thereof, in the test sample and the control sample.
 18. Themethod of claim 1, further comprising detecting at least ten genesequences selected from the group consisting of SEQ ID NO. 1 to 64 orhomologues or fragments thereof, in the test sample and the controlsample.
 19. The method of claim 1, further comprising detecting at leasttwelve gene sequences selected from the group consisting of SEQ ID NO. 1to 64 or homologues or fragments thereof, in the test sample and thecontrol sample.
 20. The method of claim 1, wherein said individual withpain is suffering from neuropathic pain.
 21. The method of claim 1,wherein detecting the level of expression in the test sample and controlsample further comprises at least one method selected from the groupconsisting of PCR of a cDNA, hybridization of a sample DNA, anddetecting one or more polypeptides encoded by said at least four genesequences or homologues or fragments thereof.
 22. A method forcharacterizing a pain status, comprising: a) providing a test samplecomprising a cell or a body fluid expressing a polynucleotide sequenceselected from the group consisting of SEQ ID NO. 1 to 64 or homologuesor fragments thereof; b) detecting expression of said polynucleotide insaid test sample; c) comparing the expression of said polynucleotide insaid test sample to expression of the same polynucleotide in a referencesample whose expression stage is known; and d) identifying a differencein the levels of expression between said test sample and said referencesample, thereby characterizing the pain status.
 23. A method foridentifying a therapeutic agent for treating pain in a subject,comprising: a) providing a test cell capable of expressing apolynucleotide sequence selected from the group consisting of SEQ ID NO:1 to 64 or homologues or fragments thereof; b) detecting expression ofsaid polynucleotide sequence in said test cell; c) contacting said testcell with the therapeutic agent; d) detecting expression of saidpolynucleotide sequence in said test cell contacted with the therapeuticagent; e) comparing the expression of said polynucleotide sequence instep (b) to the expression of said polynucleotide sequence in step (d);and f) identifying a change in expression of said polynucleotide in thepresence of the therapeutic agent, thereby identifying the therapeuticagent for treating pain.
 24. The method of claim 17, wherein said painis neuropathic pain.
 25. The method of claim 17, wherein detectingexpression of said polynucleotide further comprises at least one methodselected from the group consisting of PCR of a cDNA, hybridization of asample DNA, and detecting a polypeptide encoded by said polynucleotideor homologue or fragment thereof.
 26. A pharmaceutical composition,comprising a polynucleotide selected from the group consisting of SEQ IDNO: 1 to 64 or homologues or fragments thereof, or a polypeptide encodedby said polynucleotide.
 27. A kit comprising a reagent for detecting apolynucleotide selected from the group consisting of SEQ ID NO: 1 to 64or homologues or fragments thereof.
 28. A vector, comprising apolynucleotide selected from the group consisting of SEQ ID NO: 1 to 64or homologues or fragments thereof.
 29. A host cell, comprising apolynucleotide selected from the group consisting of SEQ ID NO: 1 to 64or homologues or fragments thereof.
 30. An antibody that selectivelybinds to a polypeptide encoded by a polynucleotide selected from thegroup consisting of SEQ ID NO: 1 to 64 or homologues or fragmentsthereof.
 31. A transgenic animal, comprising a polynucleotide selectedfrom the group consisting of SEQ ID NO: 1 to 64 or homologues orfragments thereof, wherein said polynucleotide has been altered comparedto a wild type phenotype.